Abstract In this grant we propose studies to evaluate the direct impact of Assisted Reproductive Technologies (ART) and non-In vitro Fertilization Fertility Treatments (NIFT) on the adult health of mouse offspring and to discover if epigenetic changes are present in selected tissues. These goals have wide clinical implications, since ART and NIFT are routinely used to treat patients with infertility. Currently, more than 5 million children have been conceived with ART alone. Further, embryonic and fetal developmental are stages at which the individual is vulnerable to epigenetic changes that likely contribute to disease-related outcomes. There is therefore a great need to know if preimplantation embryo manipulation induces any subtle but long-lasting effects on the health of offspring. The focus of the PAR 14-272 is at the core of the P.I.'s expertise as he has devoted most of his career to understanding how embryo stress (or non-physiological culture conditions) is associated with changes in the embryo and long-term maladaptive changes in the offspring. Data from his laboratory have demonstrated that suboptimal culture conditions affect gene expression in the mouse embryo, cause defects in placentation and lead to several postnatal metabolic deficits attributable to altered glucose homeostasis. However, multiple questions remain unanswered. For example, it is unclear how different procedures affect long-term outcome and whether epigenetic changes are present in selected tissues of ART offspring. Our laboratory is therefore uniquely positioned to generate data on the effects of different ART/NIFT procedures on these clinically relevant questions. Based on preliminary data, the primary outcomes that will be assessed are glucose homeostasis and beta cell function. The objectives of this application are to 1) understand if different ART and NIFT procedures cause diabetes in adult age and 2) discover if epigenetic changes explain alteration in glucose homeostasis observed in IVF offspring. Our central hypothesis is that manipulation of gametes, endometrium (by different hormonal stimulations) and preimplantation embryos (by culture in vitro) will result in adult onset glucose intolerance. Importantly, gametes or embryos that have undergone the most extensive manipulation will result in offspring with the most clinically relevant alteration in glucose homeostasis. Regarding expected outcomes, we will 1) determine whether offspring generated by different ART/NIFT procedures develop diabetes and 2) discover if epigenetic abnormalities are present in selected tissues of ART offspring. These datasets are expected to have an important impact in the field because of the translational value to patients affected with infertility, in addition to fundamentally advancing our understanding on how gametes and embryos respond to their environment.